Journal: Neuron

Article Title: Complement C3aR inactivation attenuates tau pathology and reverses an immune network deregulated in tauopathy models and Alzheimer’s disease

doi: 10.1016/j.neuron.2018.10.031

Figure Lengend Snippet: (A, B) C3 (A) and C3aR1 (B) mRNA levels in the parahippocampal gyrus of NCI (n=32), MCI (n=34), or AD (Dementia, n=160). Spearman’s correlation, ρ (C3)=0.36, p<2.6E-08; ρ (C3aR)=0.33, p<3.4E-07.

Article Snippet: After washing in TBS, sections or cultured cells on coverslips were blocked with TBS-T (0.4% Triton X-100) containing 2% donkey serum for 30 min, and then incubated in primary antibody diluted in blocking solution overnight at 4°C (mouse anti-GFAP (Millipore, AB_94844); rabbit anti- Iba1 (Wako, AB_839504); mouse anti-NeuN (Millipore, AB_2298772), rabbit anti-synaptophysin (Abcam, AB_2286949); rat anti-C3 (Hycult, AB_10129042); rat anti-C3aR (Hycult, AB_10130173); rabbit anti-pSTAT3 (Tyr705) (CST, AB_2491009)).

Techniques:

Journal: Neuron

Article Title: Complement C3aR inactivation attenuates tau pathology and reverses an immune network deregulated in tauopathy models and Alzheimer’s disease

doi: 10.1016/j.neuron.2018.10.031

Figure Lengend Snippet: (A)Representative GFAP and Iba1 immunostainings with DAPI in the hippocampus of 9 month-old WT, C3aR KO, PS19, and PS19/C3aR KO mice. Scale bar: 50 μM.

Article Snippet: After washing in TBS, sections or cultured cells on coverslips were blocked with TBS-T (0.4% Triton X-100) containing 2% donkey serum for 30 min, and then incubated in primary antibody diluted in blocking solution overnight at 4°C (mouse anti-GFAP (Millipore, AB_94844); rabbit anti- Iba1 (Wako, AB_839504); mouse anti-NeuN (Millipore, AB_2298772), rabbit anti-synaptophysin (Abcam, AB_2286949); rat anti-C3 (Hycult, AB_10129042); rat anti-C3aR (Hycult, AB_10130173); rabbit anti-pSTAT3 (Tyr705) (CST, AB_2491009)).

Techniques:

Journal: Neuron

Article Title: Complement C3aR inactivation attenuates tau pathology and reverses an immune network deregulated in tauopathy models and Alzheimer’s disease

doi: 10.1016/j.neuron.2018.10.031

Figure Lengend Snippet: (A) Representative Western blots of PHF1- and CP13-positive phospho-tau and total tau in PS19 and PS19/C3aR KO mouse brains at 9 months. γ-tubulin is used as a loading control.

Article Snippet: After washing in TBS, sections or cultured cells on coverslips were blocked with TBS-T (0.4% Triton X-100) containing 2% donkey serum for 30 min, and then incubated in primary antibody diluted in blocking solution overnight at 4°C (mouse anti-GFAP (Millipore, AB_94844); rabbit anti- Iba1 (Wako, AB_839504); mouse anti-NeuN (Millipore, AB_2298772), rabbit anti-synaptophysin (Abcam, AB_2286949); rat anti-C3 (Hycult, AB_10129042); rat anti-C3aR (Hycult, AB_10130173); rabbit anti-pSTAT3 (Tyr705) (CST, AB_2491009)).

Techniques: Western Blot

Journal: Neuron

Article Title: Complement C3aR inactivation attenuates tau pathology and reverses an immune network deregulated in tauopathy models and Alzheimer’s disease

doi: 10.1016/j.neuron.2018.10.031

Figure Lengend Snippet: (A) Representative synaptophysin (Syp) and Syp/NeuN co-immunostaining in area CA3 of hippocampus of WT, C3aR KO, PS19, and PS19/C3aR KO mice at 9 months. Scale bar: 50 μM.

Article Snippet: After washing in TBS, sections or cultured cells on coverslips were blocked with TBS-T (0.4% Triton X-100) containing 2% donkey serum for 30 min, and then incubated in primary antibody diluted in blocking solution overnight at 4°C (mouse anti-GFAP (Millipore, AB_94844); rabbit anti- Iba1 (Wako, AB_839504); mouse anti-NeuN (Millipore, AB_2298772), rabbit anti-synaptophysin (Abcam, AB_2286949); rat anti-C3 (Hycult, AB_10129042); rat anti-C3aR (Hycult, AB_10130173); rabbit anti-pSTAT3 (Tyr705) (CST, AB_2491009)).

Techniques: Immunostaining

Journal: Neuron

Article Title: Complement C3aR inactivation attenuates tau pathology and reverses an immune network deregulated in tauopathy models and Alzheimer’s disease

doi: 10.1016/j.neuron.2018.10.031

Figure Lengend Snippet: (A) Unsupervised clustering gene expression heatmap from RNAseq analysis of hippocampal tissues of 9 month-old WT, C3aR KO, PS19, and PS19/C3aR KO mice. 1726 DEGs were identified by comparing the PS19 vs WT group (1155 genes upregulated, red, and 571 downregulated, blue). n=4-5/genotype, adjusted p<0.01.

Article Snippet: After washing in TBS, sections or cultured cells on coverslips were blocked with TBS-T (0.4% Triton X-100) containing 2% donkey serum for 30 min, and then incubated in primary antibody diluted in blocking solution overnight at 4°C (mouse anti-GFAP (Millipore, AB_94844); rabbit anti- Iba1 (Wako, AB_839504); mouse anti-NeuN (Millipore, AB_2298772), rabbit anti-synaptophysin (Abcam, AB_2286949); rat anti-C3 (Hycult, AB_10129042); rat anti-C3aR (Hycult, AB_10130173); rabbit anti-pSTAT3 (Tyr705) (CST, AB_2491009)).

Techniques: Expressing

Journal: Neuron

Article Title: Complement C3aR inactivation attenuates tau pathology and reverses an immune network deregulated in tauopathy models and Alzheimer’s disease

doi: 10.1016/j.neuron.2018.10.031

Figure Lengend Snippet: (A) Gene-set enrichment analyses (GSEA) of the microglia sensome gene list in PS19 vs WT (left, NES=2.81, FDR<0.001) and in PS19/C3aR KO vs PS19 (right, NES= −3.0, FDR<0.001) comparisons. NES: normalized enrichment score; FDR: false discovery rate.

Article Snippet: After washing in TBS, sections or cultured cells on coverslips were blocked with TBS-T (0.4% Triton X-100) containing 2% donkey serum for 30 min, and then incubated in primary antibody diluted in blocking solution overnight at 4°C (mouse anti-GFAP (Millipore, AB_94844); rabbit anti- Iba1 (Wako, AB_839504); mouse anti-NeuN (Millipore, AB_2298772), rabbit anti-synaptophysin (Abcam, AB_2286949); rat anti-C3 (Hycult, AB_10129042); rat anti-C3aR (Hycult, AB_10130173); rabbit anti-pSTAT3 (Tyr705) (CST, AB_2491009)).

Techniques:

Journal: Neuron

Article Title: Complement C3aR inactivation attenuates tau pathology and reverses an immune network deregulated in tauopathy models and Alzheimer’s disease

doi: 10.1016/j.neuron.2018.10.031

Figure Lengend Snippet: (A, B) Total STAT3 (A) and Y705-phospho-STAT3 (B) levels in 9 month-old WT (n=10), C3aR KO (KO, n=10), PS19 (PS, n=7), and PS19/C3aR KO (PS/KO, n=7) mice measured by RPPA (mean ± s.e.m). The asterisks above PS19 are for PS19 vs WT comparison; those above PS19/C3aR KO are for PS19/C3aR KO vs PS19 comparison. Student｀s t-test. *p<0.05; **p<0.01.

Article Snippet: After washing in TBS, sections or cultured cells on coverslips were blocked with TBS-T (0.4% Triton X-100) containing 2% donkey serum for 30 min, and then incubated in primary antibody diluted in blocking solution overnight at 4°C (mouse anti-GFAP (Millipore, AB_94844); rabbit anti- Iba1 (Wako, AB_839504); mouse anti-NeuN (Millipore, AB_2298772), rabbit anti-synaptophysin (Abcam, AB_2286949); rat anti-C3 (Hycult, AB_10129042); rat anti-C3aR (Hycult, AB_10130173); rabbit anti-pSTAT3 (Tyr705) (CST, AB_2491009)).

Techniques:

Journal: Pharmaceutical Biology

Article Title: Yiguanjian decoction inhibits macrophage M1 polarization and attenuates hepatic fibrosis induced by CCl 4 /2-AAF

doi: 10.1080/13880209.2021.1961820

Figure Lengend Snippet: Primer pairs and probes used for Real-time PCR.

Article Snippet: Rat monoclonal antibody against CD68 and CD163 were obtained from Hycult Biotech (Frontstraat, UDEN, NLD).

Techniques: Sequencing

Journal: Pharmaceutical Biology

Article Title: Yiguanjian decoction inhibits macrophage M1 polarization and attenuates hepatic fibrosis induced by CCl 4 /2-AAF

doi: 10.1080/13880209.2021.1961820

Figure Lengend Snippet: YGJ inhibits liver inflammation and bile duct reaction. (a) H&E staining (×200). (b) CD68 immunostaining (×200). (c) CD163 immunostaining (×200). (d) CK19 immunostaining (×200). (e) CK19 protein bands were depicted in the immunoblot images, and (f) the densitometric quantification of the protein bands presented as a histogram ( n = 5 per group). (g) CD68, CD163, and TNF-α mRNA expressions were measured by RT-PCR and normalized to GAPDH mRNA ( n = 5 per group). (h) Serum levels of alanine aminotransferase and aspartate aminotransferase. * p < 0.05 and ** p < 0.01. N: untreated group (control); 2-AAF/CCl 4 : 2-acetylaminofluorene/carbon tetrachloride-treated group; YGJ: 2-AAF/CCl 4 + Yiguanjian decoction-treated group; SORA: 2-AAF/CCl 4 + sorafenib-treated group.

Article Snippet: Rat monoclonal antibody against CD68 and CD163 were obtained from Hycult Biotech (Frontstraat, UDEN, NLD).

Techniques: Staining, Immunostaining, Western Blot, Reverse Transcription Polymerase Chain Reaction

Journal: PLoS ONE

Article Title: The JNK Inhibitor XG-102 Protects against TNBS-Induced Colitis

doi: 10.1371/journal.pone.0030985

Figure Lengend Snippet: The proportion (%) of CD3- or CD68-positive cells from all cells within a defined vision field in the distal and medial colon from normal mice (normal, n = 5), following rectal trinitrobenzene sulfonic acid (TNBS) administration only (TNBS, n = 6) and treatment with XG-102 (100 µg/kg sc.) (n = 7). *, ** = p<0.05 and p<0.01 compared with TNBS group.

Article Snippet: Slides were incubated for 1 h at room temperature (RT) with the primary antibody against CD3 (1∶50; rabbit; Abcam, Cambridge, USA) and CD68 (1∶50; rat; Hycult Biotechnology, Beutelsbach, Germany).

Techniques:

Journal: PLoS ONE

Article Title: The JNK Inhibitor XG-102 Protects against TNBS-Induced Colitis

doi: 10.1371/journal.pone.0030985

Figure Lengend Snippet: Representative CD3 (left) and CD68 (right) immunofluorescence of the distal colon from normal mice, trinitrobenzene sulfonic acid (TNBS) administration and treatment with sc. 100 µg/kg XG-102.

Article Snippet: Slides were incubated for 1 h at room temperature (RT) with the primary antibody against CD3 (1∶50; rabbit; Abcam, Cambridge, USA) and CD68 (1∶50; rat; Hycult Biotechnology, Beutelsbach, Germany).

Techniques: Immunofluorescence